M. M. Khatun
National Institute of Biotechnology, Ganakbari, Savar, Dhaka-1349, Bangladesh
T. Tanny
National Institute of Biotechnology, Ganakbari, Savar, Dhaka-1349, Bangladesh
Abdur. M. Razzak
Pohang University of Science and Technology, Pohang 790–784, South Korea
Firoz Alam M
Department of Biotechnology and Genetic Engineering, Islamic University, Kushtia, Bangladesh
Ekhlas Uddin M
Department of Biotechnology and Genetic Engineering, Islamic University, Kushtia, Bangladesh
Ruhul Amin
Department of Biotechnology and Genetic Engineering, Islamic University, Kushtia, Bangladesh
S. Yesmin
National Institute of Biotechnology, Ganakbari, Savar, Dhaka-1349, Bangladesh
Ginger, Contamination, Surface sterilization, Rhizome buds
Plant Biotechnology Division of the National Institute of Biotechnology, Dhaka, Bangladesh
Variety and Species
Explants and nutrient medium: The experiment was conducted at the Plant Biotechnology Division of the National Institute of Biotechnology, Dhaka, Bangladesh, with the objective to evaluate the effect of different sterilants on explants of Zingiber officinale Rosc. (Ginger) in conditions of in vitro culture. BARI ada-1 (ginger) was used in this experiment. Healthy rhizomes were kept in sand for sprouting. The stored sprouted rhizomes were used to get explants. After sterilization by using different sterilants in an autoclaved beaker or conical flask, with different concentrations and duration, rhizome buds about 1 to 2 cm long were placed in a coffee jar, test tube, or conical flask on MS medium (Duchefa, The Netherlands; Murashige and Skoog, 1962) containing 3% sucrose (Merck, Germany), solidified with 0.8% agar (BDH Chemicals Ltd., England). The pH (Jenway 3520 pH Meter, Bibby Scientific Ltd., UK) of the medium was adjusted to 5.8 before autoclaving (ALP Co. Ltd., CL-40M, Japan) at 121°C and 100 kPa for 20 min and gelling with agar.
Explant sterilization: The sprouted rhizome buds were collected in beaker or conical flask and kept under running tap water prior to sterilization in the laminar airflow cabinet. For the experiment four different kinds of sterilizing agent’s viz., Mercuric (II) chloride (HgCl2), Sodium hypochlorite (NaOCl), Hydrogen peroxide (H2O2) and Bavistin, were tested for explant sterilization by vary ing their concentration and time of exposure.
Inoculation: MS basal medium supplemented with cytokinin (0.3mg/l BAP) and auxin (0.5mg/l NAA) were used for inoculation and subsequent micropropagation of Zingiber officinale Rosc. Medium was checked for the contamination before inoculation. Sterilized explants were trimmed suitably to remove sterilizing agent affected parts/brown parts. Explants were then inoculated on the appropriate medium and labeled properly. Observation were recorded regularly till to 30 days for the contamination, tissue damage and survival cultures.
Incubation: The culture were placed in culture growth room .The growth room for maintenance of in vitro cultures had 25±2°C temperature and 60 to 70% relative humidity , with a photoperiod of 16h day light and 8h dark. Illumination was provided with incandescent lamps (50 W, Philips Agro-Lite).
Statistical Analysis: Statistical analysis was done to find out the effect of different sterilizing agents its concentration and time of exposure on the asepsis of th e said plant species. For each experiment, 12 rhizome buds and conducted at least twice. The mean values and standard deviations of contamination, tissue damage and survival cultures were calculated using computer soft ware (Microsoft Office Excel Worksheet).
International Journal of Applied Biology and Pharmaceutical Technology [IJABPT], Volume-7(1):131-137, Jan-Mar-2016, ISSN: 0976-4550.
Journal